title: Protection of Enterococcus faecalis in mixed cultures with carbapenemase-producing Escherichia coli and Bacteroide fragilis: effect of the bacterial load.
abstract: BACKGROUND
  This study explores effects of pH and inoculum size on imipenem versus tigecycline activity against E. coli, B. fragilis and E. faecalis, both in individual and mixed cultures.
  
  
  METHODS
  MIC/MBCs (mg/L) of tigecycline and imipenem were 0.12/≥ 16 and 4/4 for E. coli, 0.12/0.5 and ≥ 16/≥ 16 for B. fragilis, and 0.12/≥ 16 and 2/≥ 16 for E. faecalis, respectively. Killing curves in supplemented Brucella broth were performed at pH 7 or 5.8, with two final inocula (≈ 105 or ≈ 107 cfu/ml) of each isolate (individual cultures) and with 1:1:1 mixed inocula. Tubes were 48 h incubated at 37 ºC in anaerobiosis. Final concentrations (estimated concentrations in colon) were 1.50 mg/L for tigecycline and 26.40 mg/L for imipenem, with antibiotic-free curves as controls. Experiments were performed in triplicate.
  
  
  RESULTS
  Imipenem showed inoculum effect against E.coli and B. fragilis, with reductions in initial inocula in experiments with standard inocula contrasting with increases in experiments with high inocula (both individual and mixed cultures). Against E. faecalis no inoculum effect for imipenem was observed in individual cultures, with marked reductions in initial inocula regardless inoculum size. However in mixed experiments the indirect protection of E. faecalis by the two gramnegatives resulted in bacterial regrowth. This protection was inoculum-dependant since it occurred with high but not with standard inocula. Tigecycline reduced initial inocula of the three isolates regardless culture type (individual/mixed) or experimental conditions (pH/inocula size), with lower reductions for the tolerant E. faecalis.
  
  
  CONCLUSIONS
  Carbapenemase activity was inoculum-dependant for self-protection and indirect protection of E. faecalis.
authors.affiliation: Lorenzo Aguilar, Microbiology Dept., School of Medicine, Universidad Complutense, Avda. Complutense s/n, 28040 Madrid, Spain. laguilar@med.ucm.es.
authors.forename: David
authors.initials: D
authors.lastname: Sevillano
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Aguilar
authors.forename: Luis
authors.initials: L
authors.lastname: Alou
authors.forename: María-José
authors.initials: MJ
authors.lastname: Giménez
authors.forename: Fabio
authors.initials: F
authors.lastname: Cafini
authors.forename: Natalia
authors.initials: N
authors.lastname: González
authors.forename: José
authors.initials: J
authors.lastname: Prieto
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019540:Area Under Curve; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000378:metabolism*; D001431:Bacteriological Techniques; D001441:Bacteroides fragilis / Q000187:drug effects* / Q000201:enzymology*; D003470:Culture Media; D013293:Enterococcus faecalis / Q000187:drug effects*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D015378:Imipenem / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008911:Minocycline / Q000031:analogs & derivatives / Q000494:pharmacology; D000078304:Tigecycline; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; D000078304:Tigecycline; D015378:Imipenem; D001618:beta-Lactamases; C063912:carbapenemase; D008911:Minocycline